Development of thermostable Streptokinase by recombinant DNA technology as fibrinolytic agent for different thromboembolic disorders

Background:  Streptokinase is presently utilized in clinical medicine as a therapeutic agent in the management of thromboembolic blockade. Streptokinase is an extracellular protein secreted by certain strains of beta-hemolytic streptococci. It activates plasminogen to produce plasmin, an enzyme that breaks down fibrin clots through specific lysine binding sites. Purpose of research: Production of thermostable streptokinase by recombinant DNA technology as a fibrinolytic agent for various thromboembolic diseases. The aim of this study was to improve the physicochemical properties of the streptokinase enzyme to increase its thermal stability and exhibit superior fibrinolytic activity against various thromboembolic diseases. Research type:




Screening experimental study. Our study type was to screen the thermostability and biological activity of modified streptokinase generated by bioinformatics. Methodology: Isolation of SPP-producing streptokinase was performed on blood agar. Streptococcus agalactiae was the main isolate in screening studies. In this work, streptokinase was generated by bioinformatic recombinant DNA techniques by placing cysteine-cysteine side by side in the core alpha-helix of this fibrinotyl enzyme. The streptokinase product was pre-extracted by salt precipitation with ammonium sulfate and then purified by affinity chromatography. A casein digestion method was used to detect and determine the biological activity of test enzymes compared to standard streptokinase enzymes. The expression host was Pichia pastoris SMD1168. The C-terminus is 6x histidine, the inducer is methanol, the promoter is AUG1, and PYES2-DEST52 is an expression vector. Result: This research led to improvements in the physicochemical properties of the streptokinase enzyme, making it a thermostable drug. Conclusion:Thermostable streptokinase protein produced by recombinant DNA technology, showed high efficacy as a thrombolytic agent for dissolution of various throboembolov disorders.